miR-296-3p, miR-298-5p and their downstream networks are causally involved in the higher resistance of mammalian pancreatic [alpha] cells to cytokine-induced apoptosis as compared to [beta] cells

• Published in: BMC genomics 2013-01, Vol.14
• Main Authors: Barbagallo, Davide, Piro, Salvatore, Condorelli, Angelo G, Mascali, Loriana G, Urbano, Francesca, Parrinello, Nunziatina, Monello, Adelina, Statello, Luisa, Ragusa, Marco, Rabuazzo, Agata M, Di Pietro, Cinzia, Purrello, Francesco, Purrello, Michele
• Format: Article
• Language: English
• Subjects: Apoptosis; Classification; Comparative analysis; Cytokines; Diabetes; Experiments; Gene expression; Genomes; Genomics; Methods; MicroRNA; Mobile communication systems; Pancreatic beta cells; Studies; Type 2 diabetes; Wireless communication systems
• ISSN: 1471-2164; 1471-2164
• DOI: 10.1186/1471-2164-14-62

The molecular bases of mammalian pancreatic [alpha] cells higher resistance than [beta] to proinflammatory cytokines are very poorly defined. MicroRNAs are master regulators of cell networks, but only scanty data are available on their transcriptome in these cells and its alterations in diabetes mellitus. Through high-throughput real-time PCR, we analyzed the steady state microRNA transcriptome of murine pancreatic [alpha] ([alpha]TC1-6) and [beta] ([beta]TC1) cells: their comparison demonstrated significant differences. We also characterized the alterations of [alpha]TC1-6 cells microRNA transcriptome after treatment with proinflammatory cytokines. We focused our study on two microRNAs, miR-296-3p and miR-298-5p, which were: (1) specifically expressed at steady state in [alpha]TC1-6, but not in [beta]TC1 or INS-1 cells; (2) significantly downregulated in [alpha]TC1-6 cells after treatment with cytokines in comparison to untreated controls. These microRNAs share more targets than expected by chance and were co-expressed in [alpha]TC1-6 during a 6-48 h time course treatment with cytokines. The genes encoding them are physically clustered in the murine and human genome. By exploiting specific microRNA mimics, we demonstrated that experimental upregulation of miR-296-3p and miR-298-5p raised the propensity to apoptosis of transfected and cytokine-treated [alpha]TC1-6 cells with respect to [alpha]TC1-6 cells, treated with cytokines after transfection with scramble molecules. Both microRNAs control the expression of IGF1R[beta], its downstream targets phospho-IRS-1 and phospho-ERK, and TNF[alpha]. Our computational analysis suggests that MAFB (a transcription factor exclusively expressed in pancreatic [alpha] cells within adult rodent islets of Langerhans) controls the expression of miR-296-3p and miR-298-5p. Altogether, high-throughput microRNA profiling, functional analysis with synthetic mimics and molecular characterization of modulated pathways strongly suggest that specific downregulation of miR-296-3p and miR-298-5p, coupled to upregulation of their targets as IGF1R[beta] and TNF[alpha], is a major determinant of mammalian pancreatic [alpha] cells resistance to apoptosis induction by cytokines.